Methods and articles for allergic desensitization via the oral mucosa

ABSTRACT

Compositions and methods of use for desensitizing a subject to an allergen via regions of the oral mucosa are provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.15/005,180, filed Jan. 25, 2016, which is a divisional of U.S.application Ser. No. 13/695,169, filed Feb. 1, 2013, now U.S. Pat. No.9,271,899, which is a § 371 National Stage application of PCTApplication No. PCT/US2011/034731, filed May 2, 2011, which claimspriority under 35 U.S.C. § 119(e) to U.S. Provisional Application Ser.No. 61/329,719, filed Apr. 30, 2010.

BACKGROUND

The invention relates to allergic immunotherapy targeting regions of theoral mucosa, such as those having a high dendritic to mast cell ratio,in particular targeting the vestibular mucosa. In some aspects, theinvention relates to delivery articles and dosing forms which, forexample, increase contact time and efficacy with an enhanced safetyprofile over sublingual immunotherapy.

Chronic allergic hypersensitivity disorder results from a combination ofgenetic susceptibility, failure of the immune system to properly primeduring a critical stage in infancy, and subsequent repeated exposure toa causative allergen. The sufferer develops a hyperimmune response tothe allergen that manifests clinically as allergic inflammation at thesite of contact, underpinned by antigen presenting cells, Th cells,effector cells such as mast cells, and generally IgE mediated productionof cytokines. The incidence of IgE-mediated allergic disease is spikingin industrialized Western cultures with recent estimates for totalpercent of the population afflicted in the United States running as highas about 20%. In addition to the substantial loss realized in reducedproductivity and a compromised quality of life, anaphylactic and severeasthmatic episodes may actually be life-threatening.

The clinical manifestation of a hyperallergenic immune system oftenbegins very early in life. Medical investigators have identified asyndromic profile in some individuals referred to as “atopic syndrome,”which includes a strong genetic predisposition reflected in multiplefirst and second degree familial links, coupled with an early clinicalonset of the “allergic triad” of eczema, hay fever and asthma. Acuteimmune system response manifest as, for example, food allergies andother abnormal immune responses are also very common in this population.Despite the genetic contribution to the atopic phenotype, it is alsoclear that environment plays a significant if not critical role inexpression.

Central to treatment approaches is the “hygienic hypothesis” of allergicetiology. According to this paradigm, an overly sterile environmentduring a critical stage in development of the immune system leads toaborted maturation, hypersensitivity to allergens, over production ofIgE and ultimately cytokines, and clinical symptoms of allergy. Atbirth, human infants possess a Th2 biased immune system as an artifactof the gestational period. In normal circumstances, the bias switchesafter about three to six months toward Th1. A proper Th1/Th2 balanceresults in the capacity for developing immune tolerance to environmentalallergens. The modern theory of allergy is that it reflects anabnormally persistent Th2 weighted imbalance, and immunologists havetherefore focused long-term treatment on methods for redirecting the Th2toward a Th1 response. In treatment approaches aimed at treatingindividuals with established allergic hypersensitivity, medicalclinicians use exposure to allergen to drive up the Th1 response.

Current therapy for individuals suffering from the symptoms of ahyperallergenic immune system includes medication such asantihistamines, leukotriene inhibitors and corticosteroids, and allergicimmunotherapy (AIP). Allergenic immunization has been known since theearly part of the 20^(th) century and has the goal of inducingallergen-specific tolerance in individuals exhibiting allergen-specificclinical symptoms. Subcutaneous immunotherapy, commonly referred to asSCIT, has been a treatment of choice for more than half a century. SCITis indicated where patients have a history of allergic symptoms onexposure to an offending allergen in addition to evidence of aninappropriate IgE response to that allergen. SCIT has been confirmed toinduce the desired immune response shift from a Th2 to Th1 bias and alsoto induce T(regulators) which down-regulate Th2 response. SCIT has beendemonstrated to offer significant relief from symptoms of, for example,pollen and ragweed allergy, insect venom allergy, and asthma secondaryto allergic rhinitis. Drawbacks, however, include expense,inconvenience, and the potential for induction of potentially fatalsystemic allergic reactions requiring administration of epinephrine anda need for clinical facilities and administration by highly trainedtechnicians to avoid anaphylaxis.

More recently, sublingual immunotherapy (SLIT), a therapy that evolvedas part of a maintenance adjunct with SCIT, has received considerableresearch and clinical attention. SLIT provides cheaper more convenienttherapeutic potential, possesses a strong safety profile with littlechance of a systemic response, and several recent large scale studiesconfirm the efficacy and validate the underpinning theoretical model.Generally efficacy of SLIT is based on the observation that the oralmucosa appears particularly primed in mammals for development ofallergenic tolerance and possesses specialized cells which function toachieve this. Antigen presenting cells (APC) are the initiators of theimmune response and are present in the surface of tissues incommunication with the environment. APCs capture and present allergensto T-cells, which in turn transport them to a proximal draining lymphnode for recognition and processing by effector cells including mastcells. Mast cells are the tissue based non-circulating effector cellswhich release mediators such as IgE responsible for the ultimateinflammatory response. Among APCs, dendritic cells, including thespecialized Langerhans cells, are considered the most important inducersof tolerance to allergens through their interaction with T-cells.

The mouth provides a major intake orifice for potential allergens andthe presence of dendritic cells in the oral mucosa are believed to playa role in the enhanced development of oral immune tolerance in humans.The distribution of dendritic cells, in particular Langerhans cells, inthe oral mucosa and the enhanced development of oral tolerance, takenwith the obvious advantages and conveniences associated non-invasivetherapies in general, provided the impetus to explore the oral mucosa asan alternative site for targeted allergen immunotherapy.

SLIT is typically administered as droplets or lozenges, or in tabletform intended to be held under the tongue for a few minutes and thenswallowed. Optimal dosing regimens depend on the specific targetallergen, the age of the sufferer, and the strength of the abnormalresponse sought to be ameliorated. SLIT appears to influence T andB-cell response similarly to SCIT but appears more limited. Asignificant drawback appears to be that efficacy is highly dependent onexposure time to the tissue surface, whereas the most commonly employeddosage forms are subject to rapid egress from the oral cavity andultimately digestion. Further, dosage forms designed for sustainedexposure, for example slow-dissolving lozenges held beneath the tongue,are associated with development of sublingual edema, most likely due tothe presence of mast cells located in the glands of the sublingual oralmucosa.

It was recently discovered, however, that other regions of the oralmucosa possess a dendritic cell and mast cell distribution profile thatmay be more suited for safe and effective oral immunotherapy. (See Allamet al. Allergy 2008: 63: 720-727, the entire disclosure of which isincorporated herein by this reference). According to Allam, thevestibular mucosa had the highest density of Langerhans cells(specialized dendritic cells) and a relatively low concentration of mastcells when compared with the bucca, palataum, lingua, sublingua, andgingiva. To the best of the knowledge of the present inventors, thereare no immunotherapies designed to exploit the dendritic cell and mastcell distribution profile of the oral mucosa.

There remains a need in the art for noninvasive, oral allergicimmunotherapies which retain the advantages of oral therapy but whichreduce localized safety concerns and increase efficacy over thecurrently preferred sublingual delivery region.

SUMMARY

Accordingly, the instant invention provides methods and articles formaximizing delivery of allergens to regions of the oral mucosa with adesirable relative dendritic to mast cell ratio. In particular, specificdosing and delivery methods of the invention target regions other thanthe sublingual mucosa, and certain embodiments specifically target thevestibular oral mucosa. The invention further provides methods anddelivery embodiments designed to maximize the period of contact betweenthe allergenic composition and the vestibular tissue or other targetedtissue. The dosing forms and delivery articles provided herein encouragerelatively lengthy exposure times to regions of the oral mucosa havingoptimal dendritic/mast cell distribution profiles.

One embodiment of the invention provides a flexible porous pouch forinsertion by a subject into an oral cavity of the subject and capable offixedly conforming to a vestibular mucosal surface of the oral cavity.The pouch comprises a composition formulated for extended release andthe composition comprises at least one allergen. In certain embodimentsthe pouch may be provided to the clinician or consumer as an emptyflexible pouch suitable for receiving, containing and securing thecomposition.

According to a further embodiment of the invention, methods fordecreasing sensitivity to one or more allergens and reducing symptoms ofan allergy in a subject are provided. The methods comprise providingsustained exposure of one or more allergens associated with the allergyto an oral mucosal surface having a relatively high dendritic to mastcell ratio, and in specific methods the targeted oral mucosal surface islocated substantially in a vestibular region of the oral cavity of thesubject. In specific embodiments the allergenic active may be containedin a flexible pouch according to the invention, or in a dental hygieneproduct such as toothpaste, dental cream, mouthwash or mouth spray,gels, oral strips and the like. All these dosage forms have theadvantage of exposing regions of the oral mucosa possessing a relativelyhigh number of dendritic cells, in particular the vestibular oralmucosa, to allergenic actives for a sustained period of time. Themethods avoid sustained contact with regions of the oral mucosa in closeproximity to high concentrations of mast cells associated with emptyinglymph nodes.

In some embodiments, the toothpaste, dental cream, mouthwash, ormouthspray composition induces development of immune tolerance whilepromoting oral health. The composition comprises a) toothpaste, dentalcream, mouthwash or mouthspray base ingredients; and b) at least oneallergen provided in natural form or as an extract of the allergen.Regimens suitable for optimizing the acquisition of immune tolerance tothe allergen via this delivery form reflect ordinary use of the deliveryform.

With respect to pouches, it is contemplated that allergenic active isreleased from the pouch by the action of saliva and that fulldisbursement of the active contents occurs across a prolonged periodsuch as during sleep.

In another embodiment, personalized methods for decreasing a subject'sallergic response to a specific environmental allergen are provided. Themethods comprise identifying an allergen associated with the allergicresponse, mixing an extract of the identified allergen with an organicnon-toxic filler material, filling a flexible porous pouch with themixture, wherein the filled pouch is suitable for insertion by thesubject into an oral cavity of the subject and capable of fixedlyconforming to a vestibular mucosal surface of the oral cavity, andinserting the pouch so that it fixedly conforms to the vestibularmucosal surface.

Kits are also contemplated. In one embodiment the kit comprises aplurality of flexible porous pouches comprising a composition accordingto the invention and instructions comprising a dosing schedule effectivefor desensitizing the subject to the at least one allergen. Other kitsprovide a plurality of empty flexible porous pouches along with a set ofvials containing extracts of common environmental allergens, an amountof organic non-toxic filler material; and instructions for mixingextract with the filler material and for filling the flexible porouspouch. These kits may be sold directly to consumers and may be packagedfor particular allergic disorders, such as grass allergy, allergy topets, dust mite allergy, and the like, or combinations thereof.

It is contemplated that kits may be designed on the basis of relevanceto particular geographical areas, for example.

DETAILED DESCRIPTION

All references (e.g., printed publications such as books, papers,patents, patent applications, catalogs, databases) are incorporatedherein by reference. In the event of a conflict or inconsistency, thepresent specification, as modified by any amendments thereto, shallcontrol.

Although allergic sublingual immunotherapy (SLIT) has developed a goodtherapeutic and safety reputation during its relatively shortavailability as an alternative to subcutaneous immunotherapy (SCIT), ithas recently been discovered that other regions of the oral mucosa mayprovide a superior immunogenic and safety profile.

In particular, researchers out of the University of Bonn recentlyconducted a comparative study of relevant cellular distribution acrossregions of the oral mucosa, including the bucca, gingiva, vestibula,lingua, and sublingua and reported that the vestibula had the highestdensity of Langerhans cells (specialized dendritic cells) and arelatively low level of mast cells. (Allam et al., SUMMARY, supra)Although the sublingual mucosa itself has few mast cells, they arelocated in the glands of that region and are considered responsible forthe undesirable localized side effects of itching, lingeringdiscoloration, and sublingual edema that are experienced by somesubjects undergoing SLIT regimens requiring extended exposure of theallergen specifically to the sublingual tissue.

The present invention provides dosage and delivery forms designed toresult in a distribution of allergen across the oral mucosal tissue thatcorresponds more favorably with the distribution profile of dendritic(Langerhans) cells and mast cells of the oral mucosa than dosage anddelivery forms designed specifically for maximizing exposure to thesublingual mucosa typical in SLIT.

One embodiment of the invention is directed to flexible porous pouches.The pouches are suitable for insertion by a subject into an oral cavityof the subject and capable of fixedly conforming to a vestibular mucosalsurface of the oral cavity, the pouch comprising a compositionformulated for extended release, the composition comprising at least oneallergen. (It will be appreciated that the pouch could optionally beinserted into the oral cavity of the subject by another individual.)

In some embodiments of any aspect of the invention, a composition is anutraceutical composition. A nutraceutical composition may comprise oneor more components typically found in an herbal or dietary supplement.In some embodiments, a nutraceutical composition comprises a vitamin,mineral, herb or other botanical other than tobacco (or a constituent,extract, or metabolite thereof), amino acid, fatty acid, or combinationof any of these. In some embodiments, a nutraceutical compositioncomprises one or more components isolated from a foodstuff (e.g., aplant or animal substance suitable for consumption as food fornutritional purposes by, e.g., human subjects). Typically a foodstuff isa substance normally and ordinarily acquired for consumption andconsumed by normal human subjects for nutritive and/or gustatorypurposes (as distinguished, e.g., from substances that are not normallyor ordinarily consumed but may be consumed under abnormal conditionssuch as famine or other situations in which access to customarilyconsumed foodstuffs is limited, etc.) In some embodiments of any aspectof the invention, a composition of the invention is or comprises apharmaceutical composition in accordance with the laws and/orregulations of one or more countries or regions, e.g., a country orregion in which the composition is provided to an end user, sold, and/orused. The pharmaceutical composition may be manufactured, tested,quality controlled, and/or labeled in accordance with such laws and/orregulations. In some embodiments, a pharmaceutical composition of theinvention is sold without a prescription (over-the-counter). In someembodiments, a pharmaceutical composition is provided by prescription.

Flexible pouches suitable for containing the composition are known inthe art, in particular in the context of chewing tobacco. Illustrativeexamples are disclosed in U.S. Application Serial No. 20100018540“SMOKELESS TOBACCO PRODUCTS AND PROCESSES”; U.S. Application Serial No.20070031539 “Personal caffeine delivery pouch”; U.S. Application SerialNo. 20080152695, “ORAL/BUCCAL TRANSMUCOSAL DELIVERY METHODS FORELECTROLYTE COMPOSITIONS INCLUDING XYLITOL”; U.S. Application Serial No.20080302682 “POUCH FOR TOBACCO OR TOBACCO SUBSTITUTE”; the entiredisclosures of which are incorporated herein by this reference.

In some embodiments, the composition comprises, in addition to anallergen, one or more physiologically acceptable substance(s) thatserve, e.g., as a filler material or matrix having the allergencontained therein or intermingled therewith. “Physiologically acceptablesubstance” includes substances (e.g., carriers, diluents, excipients)that do not produce an adverse or untoward reaction when administered toa mammalian subject, e.g., a human, in the amounts and at the locationsused. Numerous such substances are discussed in Remington: The Scienceand Practice of Pharmacy by University of the Sciences in Philadelphia(editor), Lippincott Williams & Wilkins; 21^(st) ed. (2005), and earliereditions thereof, and other references known to those of ordinary skillin the art. In some aspects, a polymer matrix is physically associatedwith the allergen. For example, the allergen may be entrapped, embedded,or encapsulated by the polymer matrix. A matrix can be a macroscopicstructure, which may comprise a semi-solid or viscous material, and/ormay comprise a plurality of particles (nanoparticles, microparticles). Amatrix may release the allergen by diffusion or as a result of breakdownor erosion of at least a portion of the matrix or filler material. Insome embodiments, the filler material or matrix at least in partdissolves or disintegrates over time, thereby releasing the one or moreallergens. In some embodiments, the filler material or matrix comprisesa physiologically acceptable excipient that is susceptible to cleavageby salivary enzyme(s). In some embodiments, an excipient is a polymer.In some embodiments, the polymer is a non-protein polymer, e.g., apolysaccharide. In some embodiments, the excipient is a substance thatis naturally present in one or more foods or is a generally recognizedas safe (GRAS) substance as defined by the US Food and DrugAdministration. In some embodiments, a filler material or matrixcomprises soluble or insoluble plant fiber (e.g., from a plant thatserves as a food) that resists digestive enzymes. Examples includenon-starch polysaccharides such as arabinoxylans, cellulose, and variousother plant components such as digestion-resistant dextrins, inulin, andoligosaccharides.

Extended release formulations are contemplated in order, for example, toensure sufficient exposure to effectuate desensitization in a relativelyshort time frame. In some embodiments, by prolonging the duration of atleast some exposures to the allergen, desensitization may occur withinfewer days, weeks, or months following an initial use of thecomposition, than would otherwise be the case. Formulation techniquesfor achieving extended release in a salivary environment are known inthe art. In some embodiments, an extended release formulation releasesallergen over a period of at least 5 minutes, e.g., 5-15 minutes, 15-30minutes, 30-60 minutes, 60-120 minutes, 2-4 hours, 4-8 hours, 8-12hours, etc. In particular embodiments, the extended release formulationcomprises a starch matrix incorporating the one or more allergens, saidmatrix being capable of dissolving in saliva thereby releasing the oneor more allergens into the saliva. A person of ordinary skill in the artwill be able to select formulation components which provide a desiredextended leaching of the allergenic material across the exposure timeframe.

The flexible porous pouch may be provided as a disposable single usepouch similar to pouches known for the containment of chewing tobaccoproducts. After a subject's intended use, the pouch is thrown away. Incertain embodiments, the pouch may be provided as an empty flexibleporous pouch adapted for receiving, containing and securing a dose of acomposition according to the invention. In other specific embodimentsthe flexible porous pouch may be fabricated from a material thatdissolves in saliva. In these embodiments it is contemplated that a fulldose is delivered by the point of complete dissolution.

In some embodiments, the contents of a flexible porous pouch of theinvention include, in addition to an allergen, at least one breathfreshening or flavoring ingredient. In some embodiments, use of thepouch may confer a feeling of breath freshness, well-being, and/or apleasurable gustatory sensation. In certain embodiments, use of theinventive pouch may diminish a desire for food intake.

Allergens according to the invention may include any agent whichtriggers a measurable immune response. For example, an allergen mayinclude any agent which triggers measurable production of IgE in atleast some individuals exposed to the allergen (e.g., at least someatopic individuals). In many embodiments, an allergen comprises an agentthat triggers an allergic reaction (type I hypersensitivity reaction) inat least some individuals exposed to the allergen (e.g., at least someatopic individuals). In some embodiments of the invention, the allergenis an air-borne allergen. Typically, the main route by which subject areexposed to such allergens is though inhalation. In some embodiments ofthe invention, the allergen is one to which subjects are mainly exposedby skin contact with the allergen. In some embodiments of the invention,the allergen is one to which subjects are mainly exposed by ingestingthe allergen. In some embodiments of the invention, the allergen is oneto which subjects are mainly exposed by injection.

Exemplary allergens according to the invention include allergens ofplant, animal or fungal origin. Plant allergens include pollen, sap,leaves and plant toxins, while examples of fungal allergens includepolypeptides produced by molds, Aspergillus and others. Animal allergensinclude polypeptides produced by insects, fecal allergens of dust mitesand mammals, in particular of cats, and animal keratinacious dander.Specific examples include ragweed pollen, dust mite and dust miteexcrement, animal dander and mold. Researchers have discovered that acombination of ragweed pollen and dust mite provides close to a“universal allergen” capable of affording desensitization to a widevariety of allergens. Other examples of allergens include foodallergens, various insect venoms, and a number of industrial chemicalsand pharmaceutical agents (e.g., penicillins, cephalosporins, cancerchemotherapy drugs, etc). Common sources of food allergens includepeanuts, tree nuts, eggs, milk, shellfish (e.g., shrimp, crab), fish,wheat, soy and their derivatives.

One of ordinary skill in the art will appreciate that, in general,particular allergenic molecules (e.g., particular proteins) withinallergens such as pollens, dusts, danders, molds, foods, etc, areresponsible for triggering the allergic reaction. It is common to referboth to the particular allergenic molecules (e.g., particular proteins)and the materials in which they are found as “allergens”, and thatconvention is use herein. Thus, reference to an “allergen” encompassesallergens in natural forms such as pollens, dusts, danders, molds,foods, or venoms, extracts of such natural forms of allergens, andallergenic molecules (e.g., particular proteins) that are at leastpartially purified or substantially purified or isolated from naturalsources or produced using, e.g., recombinant DNA technology. The terms“protein” and “polypeptide” are used interchangeably herein. It will beappreciated that proteins can have modifications such as glycosylation,phosphorylation, acetylation, etc., and that a protein may be a singleamino acid chain or can comprise multiple chains.

An allergen may be a modified form of a naturally occurring allergen.For example, an allergen can be chemically modified, e.g., to reduce itsallergenicity. Such modified allergens may be referred to as an“allergoid.” Allergoids may, for example, comprise allergens that havebeen treated with glutaraldehyde, formaldehyde or carbamylated. They maybe polymerized or in monomeric form.

Many allergens contain multiple distinct allergenic proteins. Numerousspecific allergenic proteins have been isolated from the naturalallergen form in which they occur and/or cDNA encoding such protein(s)has been isolated and sequenced. Amino acid sequences of numerousprotein allergens are available. Allergens may be designated as “major”and “minor” allergens. In some embodiments, a protein is considered amajor allergen if the prevalence of IgE reactivity is >50% amongindividuals sensitive to the natural form of an allergen in which theprotein occurs, with other allergens being considered “minor” allergens.In some embodiments, an allergen is a protein for which the prevalenceof IgE reactivity is >5% among individuals sensitive to the natural formof an allergen in which the protein occurs. Protein allergens of animal,plant, or fungal origin are usually named using a systematicnomenclature developed by the World Health Organization andInternational Union of Immunological Societies (WHO/IUIS) AllergenNomenclature Sub-committee under the auspices of the WHO and IUIS (see,e.g., Lockey, R F and Ledford, DK (eds.) “Allergens and AllergenImmunotherapy” 4th ed. 2008. Informa Healthcare, New York, incorporatedby reference herein, e.g., Chapman MD. Allergen Nomenclature. Chapter 3(pp. 47-58) therein. According to this nomenclature, the first threeletters of the genus are followed by the first letter of the species andthen a numeral. For example: Phl p 5 is a major allergen of Phleumpretense (timothy grass) pollen. The WHO/IUIS Allergen NomenclatureSub-committee maintains an allergen database (WHO/IUIS AllergenDatabase) that contains numerous approved and officially recognizedallergens. The database, which can be accessed on the website availableat www.allergen.org, is searchable by allergen name and allergen source(common or scientific name). According to the WHO/IUIS nomenclature,isoallergens are defined as allergens from a single species, sharingsimilar molecular size, identical biological function, and greater than67% amino acid sequence identity (Chapman MD, supra). It will beappreciated that multiple isoforms (variants that differ in amino acidsequence) of many allergens are found in nature. Isoallergens andisoforms are denoted by the addition of four numeral suffixes to theallergen name. The first two numerals distinguish between isoallergensand the last two between isoforms. Such two or four numeral suffixeswill generally be omitted herein, but it should be understood that thevarious allergen isoallergens and isoforms known in the art are includedwithin the scope of allergens of use in embodiments of the variousaspects of the instant invention. One of ordinary skill in the art willreadily be able to obtain amino acid sequences for numerous proteinallergens of plant, animal, or fungal origin, among others, as well assequence of nucleic acids encoding such allergens, using publiclyavailable information. For example, the WHO/IUIS Allergen Databaseprovides UniProt accession number for numerous protein allergens andGenbank accession number for nucleic acids encoding them. SDAP(Structural Database of Allergenic Proteins) is a Web server (availableat the University of Texas Medical Branch website available atfermi.utmb.edu/SDAP) that, among other things, allows the user toretrieve information for allergens (e.g., sequence information) from themost common protein sequence and structure databases (SwissProt, PIR,NCBI, PDB). The US National Center for Biotechnology Information (NCBI)databases (available at www.ncbi.nlm.nih.gov) such as Genbank provideinformation regarding amino acid sequences of numerous protein allergensand nucleic acids that encode them. UniProt accession numbers (acc. no.)for various allergens of interest are provided herein for illustrativepurposes. The afore-mentioned databases are incorporated herein byreference, e.g., allergen names, accession numbers, and sequences, areincorporated herein by reference. One of ordinary skill in the art canreadily identify sequences of other allergens. As noted above, one ofordinary skill in the art will appreciate that isoallergens and isoformsof many of these allergens exist. One of skill in the art will furtherappreciate that variations in one or more nucleotides of the nucleicacids encoding a particular protein may exist among individuals of agiven species due to natural allelic variation. As a result of thedegeneracy of the genetic code, many of these variations do not resultin changes in amino acid sequence. One of ordinary skill in the art willfurther appreciate that an allergen protein may be synthesized as aprecursor protein that contains one or more portions not found in themature form. A mature allergen protein may have been processedintracellulary or extracellularly so as to remove one or more portion(s)of the preprotein. For example, a signal peptide may be removed, or apolypeptide chain may be cleaved to form two or more chains, optionallyremoving a portion of the precursor protein.

Plant pollens are major sources of airborne allergy throughout manyareas of the world. In some embodiments of the invention, an allergencomprises grass pollen. Grasses, as used herein, include members of thefamily Poaceae (sometimes termed “true grasses”), rushes (Juncaceae) andsedges (Cyperaceae). Grasses are distributed widely throughout manyregions of the world, with different species having variable importancein different geographical areas. For example, grass species common in atleast some regions of Europe and/or the US include Dactylis glomerata(orchard grass), Poa pratensis (Kentucky bluegrass), Lolium perenne(ryegrass), Anthoxantum odoratum (sweet vernal), Phleum pratense(timothy), Festuca eliator (meadow fescue), Agrostis alba (redtop), andCynodon dactylon (Bermuda grass). Grass allergens include, e.g., Poa a 1(UniProt acc. no. Q9ZP03) and Poa p 5 (UniProt acc. no. Q9 FPR0). Insome embodiments of the invention, an allergen is from a grass withinthe Dactylis, Poa, Lolium, Anthoxantum, Phleum, Festuca, Agrostis, orCynodon genus, e.g., any of the afore-mentioned species. For example, anallergen can comprise a Poa a, Poa p, or Phl p protein.

In some embodiments of the invention, an allergen is pollen (or anextract or component thereof) of a tree or shrub that is a member of theCupressaceae family. It should be noted that the Cupressaceae (cypress)family includes a number of species whose common name includes the word“cedar.” In some embodiments, the allergen is pollen from a species inthe subfamily Cupressoideae, e.g., a member of the genus Chamaecyparisor Juniperus (“juniper”). In some embodiments, the allergen is pollenfrom Cryptomeria japonica (family Cupressaceae, subfamily Taxodioidea),commonly referred to as Sugi or Japanese cedar. Japanese cedar pollen isthe major cause of pollinosis in Japan. Approximately 15% of theJapanese population was affected by Japanese cedar pollinosis in 2002(Okuda M., Ann Allergy Asthma Immunol, 91: 288-96, 2003), and theprevalence has reportedly increased to an estimated 26.5% in 2008 (seeOkubo, K., Allergol Int., 57(3):265-75 2008, incorporated by reference).Many patients with cedar pollinosis have also been sensitized toChamaecyparis obtusa pollen (Japanese cypress, hinoki cypress orhinoki), which disperses after Japanese cedar pollen. In theseindividuals, symptoms of Japenese cedar pollinosis are frequentlyfollowed by those of cypress pollinosis, often resulting in asymptomatic period lasting for about 4 months (e.g., from February toMay). In some embodiments, the invention provides compositions andmethods of use in desensitizing individuals who suffer from Japanesecedar and/or Japanese cypress pollinosis. Cry j 1 (UniProt acc. no.P18632) and Cry j 2 (UniProt acc. no. P43212) are major allergens ofCryptomeria japonica pollen. See, e.g., Yasueda H, et al., J AllergyClin Immunol., 71(1 Pt 1):77-86, 1983; Sakaguchi M, et al. Allergy,45:309-312, 1990, for discussion. cDNAs encoding these allergen proteinshave been cloned and sequenced. See, e.g., Sone T, et al., BiochemBiophys Res Commun. 199:619-625, 1994; Komiyama N, et al. BiochemBiophys Res Commun. 201:1021-1028, 1994; Namba M, et al., FEBS Lett.353:124-128, 1994; and PCT/US1992/005661 (WO1993001213—ALLERGENICPROTEINS AND PEPTIDES FROM JAPANESE CEDAR POLLEN). Cry j 3 has beenidentified, and sequences are available (see, e.g., Futamura N, et al.Biosci Biotechnol Biochem. 66(11):2495-500, 2002; Futamura N, et al.Tree Physiol. 26:51-62, 2006). Other allergens, e.g., Cry j 4, Cry j 5,Cry j 6 have been identified as well (Matsumura D, et al., Biol PharmBull. 29(6):1162-6; 2006). In some embodiments, an allergen comprises aCry j protein, e.g., Cry j 1, Cry j 2, Cry j 3, Cry j 4, Cry j 5, Cry j6.

Cha o 1 (UniProt acc. no. Q96385) and Cha o 2 (UniProt acc. no. Q7M1E7)are major allergens of Japanese cypress, cDNAs for which have beencloned and sequenced (Suzuki M, et al.., Mol Immunol. 33(4-5):451-60,1996; Mori T, et al, Biochem Biophys Res Commun. 263(1):166-71, 1999).In some embodiments an allergen comprises a Cha o protein, e.g., Cha o1, Cha o 2.

Ashe juniper (Juniperus ashei, family Cupressaceae, sometimes calledmountain cedar) and Arizona cypress (Cupressus arizonica, familyCupressaceae) pollens cause seasonal allergic rhinitis in certain partsof the US and Northern Mexico while Italian cypresses (Cupressussemperverins, family Cupressaceae) cause pollinosis in the Mediterraneanregion (e.g., France, Italy, Israel). In some embodiments, the inventionprovides compositions and methods of use in desensitizing individualswho suffer from allergy to one or more such pollens. Jun a 1 and Jun a 2are major allegens of juniper pollen. See, e.g., Midoro-Horiuti T, JAllergy Clin Immunol. 104(3 Pt 1):608-12, 1999; Midoro-Horiuti T, JAllergy Clin Immunol. 104(3 Pt 1):613-7, 1999; Yokoyama M, BiochemBiophys Res Commun. 275(1):195-202, 2000). The amino acid sequence ofJun a 1 (UniProt acc. no. P81294) shows significant identity with thoseof Cry j 1 and Cha o 1, respectively. The amino acid sequence of Jun a 2(UniProt acc. no. Q9FY19) shows about 70% and 82% identity with those ofCry j 2 and Cha o 2, respectively. In some embodiments an allergencomprises a Jun a protein, e.g., Jun a 1, Jun a 2.

The Betulaceae, or birch family, includes six genera of deciduousnut-bearing trees and shrubs, including the birches (genus Betula),alders (genus Alnus), hazels (genus Corylus), hornbeams andhop-hornbeams. In some embodiments of the invention, the allergencomprises pollen (or an extract or component thereof) of a member of thebirch family. In some embodiments the pollen is from a member of thesubfamily Betuloideae. In some embodiments, the pollen is from genusBetula, e.g., Betula verrucosa. Birch pollen allergens include, e.g.,Bet v 1, Bet v 2, Bet v 3, Bet v 4, Bet v 5, Bet v 6, and Bet v 7. Insome embodiments, the pollen is from genus Alnus, e.g., Alnus glutinosa.Alder pollen allergens include, e.g., Aln g 1 and Aln g 4. In someembodiments, the pollen is from genus Corylus, e.g., Corylus avellana.In some embodiments, an allergen comprises a Bet v protein.

Various other plants that are significant causes of allergy belong tothe families Asteraceae, Amaranthaceae, Urticaceae, Euphorbiaceae, andPlantaginaceae. In some embodiments of the invention, a compositioncomprises a pollen (or an extract or component thereof) from a plant ofthe Asteraceae, Amaranthaceae, Urticaceae, Euphorbiaceae, orPlantaginaceae family. Examples of such plants include ragweed,cocklebur, marsh elder, mugwort, feverfew, pellitory, goosefoot,plantain, and Russian thistle. Ragweeds (Ambrosia species), for example,are a genus of flowering plants from the sunflower family (Asteraceae)and represent a highly significant cause of allergy in North Americathat is becoming increasingly important in Europe. Four major familiesof proteins may represent the major cause of allergic reactions topollens of such plants: the ragweed Amb a 1 family of pectate lyases(e.g., UniProt accession numbers P27759, P27760, P27761, P27762 fromAmbrosia artemisiifoha (short ragweed)); the defensin-like Art v 1family (e.g., from mugwort and feverfew, e.g., UniProt acc. no. Q84ZX5from Artemisia vulgaris (mugwort)); the Ole e 1-like allergens, Pla 1 1from plantain, and Che a 1 from goosefoot, and the nonspecific lipidtransfer proteins Par j 1 and Par j 2 from pellitory (Gadermaier G, etal. Curr Allergy Asthma Rep. 4(5):391-400, 2004)). Amb a 1 was among thefirst of these allergens for which cDNA was cloned and sequenced. See,e.g., PCT/US1990/001310 (WO/1990/011293—ALLERGENIC PROTEINS FROM RAGWEEDAND USES THEREFOR). In some embodiments of the invention, an allergencomprises an Amb a, Art v, Ole e-like, Pla 1, Par j protein, orcombination thereof. Mixtures of pollens from such plants (and extractsand components thereof) are contemplated. Plant allergens of naturalrubber latex derived from a variety of different plant species (e.g.,Hevea, such as Hevea brasiliensis) are contemplated.

Dust mites are significant sources of allergy in many areas of theworld. Allergens are found in dust mite feces and the mite body. Dustmite species of significant importance include, for example,Dermatophagoides farinae, D. pteronyssinus, and Tyrophagusputrescentiae. House dust mite allergens include, for example, Der p1(UniProt acc. no. P08176), Der p 2 (UniProt acc. no. P49278), Der p 3(UniProt acc. no. P39675), and Der p 4 from D. pteronyssinus and Der f 1(UniProt acc. no. P16311), Der f 2 (UniProt acc. no. Q00855), and Der f3 (UniProt acc. no. P49275) from D. farinae. In some embodiments of theinvention an allergen comprises a Def p, Der f, or Tyr p protein.

Animal allergens occur, for example, in dander, feathers, hair, saliva,and excretions (e.g., urine). Domesticated animals such as cats (Felisdomesticus) and dogs (Canis lupus familiaris) are common sources ofallergy. Fel d 1 (UniProt acc. no. P30438 (chain 1); UniProt acc. no.P30440 (chain 2)), Fel d 3, and Fel d 4 are major cat allergens. Can f 1(UniProt acc. no. 018873) and Can f 2 (UniProt acc. no. 018874) aremajor dog allergens. In some embodiments an allergen comprises a Can forFel d allergen. Rodents such as mice (e.g., Mus musculus), rats (e.g.,Rattus norvegicus), and rabbits (e.g., European rabbit (Oryctolaguscuniculus)) are common sources of allergy. Identified allergens include,e.g., Mus m 1, Rat n 1, and Ory c 1, in these species, respectively.Individuals may, for example, encounter such animals as pets, as pests,or in an occupational context (e.g., as laboratory animals). In someembodiments an allergen comprises a Mus m, Rat n, or Ory c protein. Farmanimals such as horses, cows, sheep, and goats are also causes ofallergy, and in some embodiments an allergen from such animal is presentin an inventive composition.

Insects and insect venoms are notable sources of allergens. Cockroachallergens are significant causes of allergy in many areas of the world.Cockroach species include, for example, Blattella germanica (Germancockroach) and Periplaneta americana (American cockroach), and Blattaorientalis (Oriental cockroach) Cockroach allergens include, forexample, Bla g 1, Bla g 2, Bla g 5, Bla g 5, Bla g 6, Bla g 7, and Bla g8 (from B. germanica) and Per a 1, Per a 3, Per a 6, Per a 7, Per a 9,and Per a 10 (from P. Americana). In some embodiments an allergencomprises a Bla g, Per a, or Bla o allergen. Ant, moths, fleas, flies(e.g., house fly, horse fly, mayfly), and mosquitos are also sources ofallergens. In some embodiments an allergen is a cockroach, ant, moth,flea, fly, or mosquito protein.

Insect venoms, (e.g., from insects of the order Hymenoptera, e.g., bees,hornets, or wasps) that are potential causes of severe allergicreactions include venoms from European Hornet (Vespa crabro), Honey Bee(Apis mellifera), Hornet (Dolichovespula spp.), Paper Wasp (Polistesspp.), Yellow Jacket (Vespula spp.), White (Bald)-Faced Hornet(Dolichovespula maculata), Yellow Hornet (Dolichovespula arenaria). Insome embodiments an allergen is a venom (or extract or componentthereof) of a bee, wasp, or hornet. For example, an allergen cancomprise an Api, Dol, or Ves protein.

Fungi (e.g., fungal spores or fragments (e.g., hyphal fragments)) aresignificant sources of allergy. Alternaria (e.g., Alternaria alternata(Alternaria rot fungus)), Cladosporium (e.g., Cladosporium herbarum,Cladosporium cladosporioides), Aspergillus (e.g., Aspergillus fumigatus,Aspergillus niger), Fusarium, Penicillium are exemplary allergenic fungiof interest. In some embodiments, an allergen comprises a protein foundin or produced by Alternaria, Cladosporium, Aspergillus, Fusarium,Penicillium, or other fungus. For example, an allergen can comprise anAlt a, Asp a, Asp n, Cla or Pen protein.

Methods of obtaining allergens are well known in the art. For example,pollens can be collected from the respective plants, which may becultivated or in the wild. Fungal extracts can be produced from pureculture mycelial mats or allergens can be isolated from culture medium.Rusts and smuts can be obtained from natural growths. Epithelialextracts can be produced from the hide, hair, or feathers containing thenatural dander, or from separated dander. Insect and mite extracts canbe produced from the whole body of the insects or mite, respectively. Inthe case of insect venoms, venom or venom-containing organs can beisolated or a whole body extract can be used. House dust can be madefrom various dusts ordinarily found in the home (e.g., upholstery dust,mattress dust, or general dust accumulating on surfaces). Other dusts(e.g., grain dust, wood dust, cotton dust) can be collected from theappropriate location. Food extracts can be prepared from the edibleportions of the respective foods, e.g., freshly obtained foods.

Methods suitable for allergen processing, e.g., production of allergenextracts, purification of allergen molecules, etc., are well known inthe art. Very briefly, source allergen material (e.g., pollen, insect,dander) can be subjected initially to pulverization, drying, defatting(by extraction using organic solvent), or other steps as appropriate forthe particular allergen. Centrifugation can be used, e.g., to separatesolid or particulate matter. Resulting material can be incubated in anaqueous medium (e.g., water or suitable buffered solution, e.g.,ammonium bicarbonate, phosphate buffered saline, etc.) for a suitableperiod of time to at least partly solubilize proteins. Crude extract canbe processed using, e.g., dialysis, filtration, fractionation,chromatography, etc. In some embodiments, one or more steps is performedto at least partly remove low molecular weight components, concentratethe extract, etc. Extracts can be sterilized, e.g., using filtrationand/or irradiation. Other processing steps can be applied as known inthe art. Numerous specific protocols are available.

Extracts of allergens specifically processed for safe use in humanimmunotherapy are available commercially. For example, GREERLaboratories Inc. Allergy and Immunotherapy division publishes abrochure entitled, “Human Allergy Products and Services” availableon-line at the company website currently atwww.greerlabs.com/files/catalogs/HumanAllergyCatalog.pdf. GREER alsopublishes a brochure entitled “Source Materials Products and Services”available online at the company website currently atwww.greerlabs.com/files/catalogs/SourceMaterialsCatalog.pdf, whichdetails available allergens that can be used as raw materials forproduction of allergen extracts or more highly purified allergen proteinpreparations. Both publications are incorporated herein by reference.Other commercial suppliers of allergens and/or allergen extracts includeALK Abello, Inc., Allermed Labs, and HollisterStier. An extensive listof allergen extracts is found in Remington, supra. Allergen extractstypically contain multiple proteins, e.g., multiple allergenic proteins,present in the natural form of the allergen. Extracts can be preparedfrom, e.g., pollens (e.g., of trees, shrubs, grasses, other plants suchas those often termed “weeds”), animal epithelia, feathers, fungalmycelia or spores, smuts, mites, insects, insect venoms, foods, dusts,etc. In some embodiments, an extract is prepared essentially from asingle natural allergen (e.g., obtained from a single species of plant,animal, insect, fungus, etc.). Mixtures are contemplated. In someembodiments an extract is derived from multiple different plant pollens(e.g., weed mixture, tree mixture, grass mixture), multiple differentfungi or smuts, multiple different insect venoms, multiple differentanimal epithelia, etc. Fungal extracts can be prepared from myceliaand/or spores (e.g., Alternaria, Cladosporium) and/or from culturefiltrate material (e.g., Aspergillus). In some embodiments, one or moreallergen protein(s) is further purified, e.g., from an extractcomprising multiple proteins. One of ordinary skill in the art willreadily be able to purify allergen protein(s) of interest using methodsknown in the art for protein purification. See, e.g., Cutler, P. (ed.)Protein Purification Protocols, Methods in Molecular Biology, Volume244, 2004; Simpson, R J, et al., Basic Methods in Protein Purificationand Analysis: A Laboratory Manual Cold Spring Harbor Laboratory Press,2008; Richard R Burgess and Murray P. Deutscher (eds.) Methods inEnzymology: Guide to Protein Purification, 2^(nd) ed., Academic Press,2009. Purification can entail chromatographic methods (e.g., based onsize, hydrophobicity, affinity, etc.), immunological methods,electrophoretic methods, etc. Specific protocols for preparing variousat least partially purified allergen proteins are available. In someembodiments, an extract or at least partially purified proteinpreparation comprises at least 70%, 80%, 90%, 95% or more protein byweight. In some embodiments, a protein is considered pure when it isremoved from substantially all other compounds or entities other than asolvent and any ions contained in the solvent, e.g., the proteinconstitutes at least about 90%, e.g., at least 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99%, or greater than 99% of the dry weight of thecomposition or on a weight per volume basis (excluding the solvent andions). In some embodiments, a particular allergen protein of interest isconsidered pure if it constitutes at least about 90%, e.g., at least91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater than 99% of theprotein content in the protein preparation by dry weight. Methods forassessing purity are known in the art and include, e.g., chromatographicmethods, immunological methods, electrophoretic methods, massspectrometry, etc. Any of the polypeptides described herein may bepurified, in various embodiments. An extract or purified proteinpreparation can be provided in various formats. For example, an extractor purified protein preparation dried, e.g., lyophilized, or provided inaqueous medium, optionally comprising a protein stabilizing agent suchas glycerin, a preservative, etc.

Mixtures of individual allergen proteins are contemplated. Allergenprotein mixtures can comprise allergen proteins from the same species orfrom multiple different species, which may be in the same or differentgenera, subfamily, family, etc. In some embodiments, an allergen proteinencoded by a gene homologous to that which encodes a particular allergenprotein of interest can be used. For example, orthologous genes, i.e.,genes in different species that are similar to each other because theyoriginated by vertical descent from a single gene of the last commonancestor, can be used.

Also contemplated as within the scope of the invention are nutraceuticalfood additives, for example plant extracts, with demonstratedimmunosuppressive character and/or confirmed efficacy in diminishingallergic response.

In some embodiments, an allergen comprises a recombinantly producedprotein. Methods for producing proteins using recombinant DNA technologyare well known in the art and are described in standard references suchas Ausubel, F., et al., (eds.), Current Protocols in Molecular Biology,Current Protocols in Immunology, Current Protocols in Protein Science,and Current Protocols in Cell Biology, all John Wiley & Sons, N.Y.,editions as of 2008; Sambrook, Russell, and Sambrook, Molecular Cloning:A Laboratory Manual, 3^(rd) ed., Cold Spring Harbor Laboratory Press,Cold Spring Harbor, 2001; Harlow, E. and Lane, D., Antibodies—ALaboratory Manual, Cold Spring Harbor Laboratory Press, Cold SpringHarbor, 1988; Burns, R., Immunochemical Protocols (Methods in MolecularBiology) Humana Press; 3rd ed., 2005, all of which are incorporatedherein by reference. Any suitable vectors, e.g., plasmids, viruses(e.g., DNA or RNA viruses), cosmids, etc., can be used to introduce anucleic acid that encodes an allergen protein into a host cell, invarious embodiments. One of ordinary skill in the art would appreciatethat due to the degeneracy of the genetic code, any of a wide variety ofnucleic acid sequences can encode a protein of interest (e.g., anallergen) and can accordingly be used in various embodiments of theinvention relating to recombinant production of allergens. In someembodiments, a nucleic acid sequence is codon optimized for productionof the protein in a host cell of interest. Any suitable expressionsystem can be used. Various host cells, e.g., bacterial, fungal, insect,vertebrate (e.g., mammal), can be used in various embodiments. In someembodiments a host cell is selected based at least in part on theallergen. For example, in some embodiments a plant allergen can beproduced in plant cells; a vertebrate allergen can be produced invertebrate cells; a fungal allergen can be produced in fungal cells. Anallergen could be produced using a transgenic approach, e.g., atransgenic plant. In some embodiments, the sequence of a recombinantlyproduced allergen comprises a fragment or variant of the sequence of anaturally occurring allergen protein. For example, a fragment may be acontinuous sequence consisting of at least 20%, 30%, 40%, 50%, 60%, 70%,80%, 90%, or more of the full length naturally occurring allergenprotein. A variant has one or more amino acid substitutions, deletions,or additions (e.g., insertions) as compared with a naturally occurringallergen protein. For example, a variant may comprise a sequence atleast 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more identical to anaturally occurring allergen protein over a window of at least 50%, 60%,70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more of the naturallyoccurring antigen, allowing the introduction of gaps to maximizeidentity. Percent identity can be calculated with the use of a varietyof computer programs known in the art. For example, computer programssuch as BLAST2, BLASTN, BLASTP, Gapped BLAST, etc., generate alignmentsand provide percent identity between a sequence of interest andsequences in any of a variety of public databases. The algorithm ofKarlin and Altschul (Karlin and Altschul, Proc. Natl. Acad. Sci. USA87:22264-2268, 1990) modified as in Karlin and Altschul, Proc. Natl.Acad. Sci. USA 90:5873-5877, 1993 is incorporated into the NBLAST and)(BLAST programs of Altschul et al. (Altschul, et al., J. Mol. Biol.215:403-410, 1990). To obtain gapped alignments for comparison purposes,Gapped BLAST is utilized as described in Altschul et al. (Altschul, etal. Nucleic Acids Res. 25: 3389-3402, 1997).

When utilizing BLAST and Gapped BLAST programs, the default parametersof the respective programs are used. A PAM250 or BLOSUM62 matrix may beused. See the Web site having URL www.ncbi.nlm.nih.gov for theseprograms. In a specific embodiment, percent identity is calculated usingBLAST2 with default parameters. In some embodiments, a variant has up toabout 1%, 5%, 10%, 20%, or 30% amino acid substitutions, insertions, ordeletions. In some embodiments, a variant has between 1 and 10 aminoacid substitutions, insertions, or deletions.

In some embodiments, an allergen may be a fusion protein comprising atleast a portion of an allergen protein comprising at least oneallergenic epitope and a heterologous polypeptide. In some embodiments,the fusion protein comprises a tag, e.g., an epitope tag, of use inpurifying the protein. In some embodiments, an allergen, e.g., a fusionprotein, is expressed in a plant, e.g., in leaves or seeds. Such leavesor seeds can be used to prepare a composition of this invention. Forexample, a nucleic acid encoding the allergen can be expressed undercontrol of an appropriate promoter to achieve expression in the plant ora portion thereof. A plant can be a transgenic plant or the protein canbe transiently expressed, e.g., using a viral vector. In someembodiments a fusion protein comprises an allergenic epitope (e.g., anepitope of Japanese cedar pollen or other allergenic pollen) fused withat least a portion of a seed storage protein such as glycinin. A fusionprotein could comprise multiple different allergen proteins or portionsthereof. Methods such as solid phase peptide synthesis or proteinligation could be used to synthesize polypeptides in some embodiments,particularly if relatively short.

In some embodiments, a polypeptide that comprises one or moreepitope(s), e.g., epitope(s) recognized by human T-cells derived from anaturally occurring allergen is used as an allergen in the instantinvention. In some embodiments, the polypeptide comprises human T-cellepitope(s) derived from at least two different allergens, e.g., 2, 3, 4,5, 6, or more different allergens. In some embodiments, the total numberof T-cell epitopes is between 2 and 10. The epitopes may be separatedfrom each other by 1 or more amino acids that serve as a spacer. Aspacer can be, e.g., 1, 2, 3, 4, or 5 amino acids long, up to about 25amino acids. In general, any amino acids can be used as spacers. In somenon-limiting embodiments, relatively small amino acids such as Gly, Ala,Ser are used, but other amino acids could be used. For example,Cry-consensus peptide is a polypeptide containing six major human T-cellepitopes derived from both Cry j 1 and Cry j 2 (Tsunematsu M, et al.Allergology International. 56(4):465-72, 2007). Its sequence is:MKVTVAFNQFGPN-RR-VFIKRVSNVIIHG-RR-IDIFASKNFHLQKNTIGTG-RR-WKNNRIWLQFAKLTGFTLMG-RR-LKMPMYIAGYKTFDG-RR-VDGIIAAYQNPASWK(SEQ ID NO: 1). The T-cell epitopes are underlined. Epitopes 1, 2, and 5(starting from the left) are Cry j 1 epitopes. Epitopes 3, 4, and 6 areCry j 2 epitopes. T cell epitopes of a variety of allergens have beenidentified and could be used in embodiments of the invention. An epitopecan be a linear epitope or a conformational epitope.

According to specific embodiments, the composition, comprises allergensselected from the group consisting of pollen, dust mite, mold and animaldander. Personalized regimens are contemplated wherein the subject orthe subject's medical advisor selects allergens based on specificclinical symptoms wherein the allergen is relevant to the subject'senvironment, such as specific pet dander, geographically circumscribedpollens, or barnyard dander and the like. In personalized methods thesubject exhibits clinical symptoms of an allergy and the one or moreallergens are selected based on association with the allergy. In veryspecific embodiments based on a “universal” allergen approach, theallergen consists of ragweed pollen and dust mite, although thecomposition may comprise other allergens as well. In some embodiments,an allergen consists of a pollen allergen and an insect allergen. Insome embodiments, an allergen consists of a pollen allergen and a miteallergen.

In some embodiments, an allergen extract is provided in a liquid form.For example, an allergen extract can be provided in water, glycerin, ora combination thereof. In some embodiments, an allergen extract isprovided in substantially dry form, e.g., as a powder, e.g., inlyophilized form. In some embodiments, an allergen is provided in tabletform, wherein the tablet is inside the pouch, and the pouch optionallyfurther contains one or more substances that serve as a filler material.In at least some of these embodiments, an allergen-containing tabletsuch as those used in SLIT (or contents thereof) can be employed. Thetablet can contain allergen extract in lyophilized form. In someembodiments the tablet is a rapidly disintegrating tablet. In someembodiments, a five-grass pollen SLIT tablet (Stallergènes SA, France)can be used. The extracts are from pollen of perennial rye grass (Loliumperenne), meadow grass (Poa pratensis), timothy grass (Phleum pratense),cocksfoot (Dactylis glomerata) and sweet vernal grass (Anthoxanthumodoratum). and are sold under the name Oralair®. In some embodiments, aGrasax® tablet is used (ALK Grass tablet, ALK-Abelló A/S, Hørsholm,Denmark). The active substance in Grasax is a standardised allergenextract of grass pollen from timothy (Phleum pratense). Otheringredients in Grasax are gelatine (fish source), mannitol, and sodiumhydroxide. Of course other excipients could be used as known in the artof tablet formulation. The tablet can be provided within a flexibleporous pouch, optionally together with a filler material. The tablet maybe at least in part surrounded by the filler material. In someembodiments, the allergen diffuses through the filler material to reachthe vestibular, buccal, and/or gingival mucosa. The filler material mayprovide for an increased contact area of the allergen with the oralmucosa as compared with the area of the tablet.

One of ordinary skill in the art will appreciate that the amount ofallergen(s) used in a composition of the invention (e.g., compositionfor inclusion in a flexible porous pouch or a dental hygiene composition(as discussed below)) can vary based on a number of factors such as, forexample, the particular allergen(s) and the potency of the allergenpreparation used. Allergens can be detected and/or quantified usingmethods known in the art, and such methods can be used to characterizeallergen preparations (e.g., allergen extracts), e.g., with respect toallergen content and/or potency. For example, immunological methods suchas immunoblotting or ELISA assays using, e.g., an appropriate monoclonalantibody, or other types of binding assays for detecting and/orquantifying proteins known in the art are applicable to many allergensof interest herein. Mass spectrometry could be used. In vivo or in vitrobioassays may be used to quantify biological activity. For example, skinprick tests can be performed on individuals known to be allergic to aparticular allergen, and one or more indicators of allergic response(e.g., wheal area) or allergen-specific IgE production, can be measured.Skin prick tests can be standardized, e.g., using a specific deliverydevice, technique, and specified amount of allergen preparation. Invitro bioassays include measuring mediator release (e.g., histamine,cytokines, or lipid mediators) from appropriate cells, e.g., sensitizedcells. An in-house reference standard can be produced (e.g., based on invivo bioassays) and in vitro tests can be used to compare the potency ofsubsequent batches of allergen with the in-house reference and potencycan be assigned as arbitrary units. A variety of units are in use toquantify allergen content and/or potency of allergen preparations. See,e.g., Remington, supra, and Lockey, R F and Ledford, DK, supra. Forexample, allergen content of allergen preparations can be expressed asweight-to-volume or Protein Nitrogen Units. In the US, standardizedextracts are typically labeled as Allergy Units (AU)/ml or BioequivalentAllergy Units (BAU)/ml. BAU is a standard established by the FDA, whichprovides a variety of different reference extracts that can be used toestablish potency. In Japan, allergen content of allergen preparationsis often expressed using Japanese Allergy Units (JAU) measured by ELISA.In Europe, a number of country-specific or company-specific standardsare in use. For example, in-house reference standards can be establishedand used to quantify the strength of subsequently produced batches ofallergen preparation. For example, an in-house referenceindex-of-reactivity (IR) can be used, wherein, for example, 100 IR/ml isdefined as the concentration eliciting, by means of skin prick testing,a geometric mean wheal size of 7 mm in diameter in 30 patients sensitiveto the corresponding allergen. If desired, any one or more of suchapproaches can be employed with regard to the allergen preparations ofuse in the instant invention.

In some embodiments, a composition of the invention (e.g., a compositionfor inclusion in a flexible porous pouch or a dental hygiene product orchewing gum) can be formulated so that a typical use provides between 1pg to 15 mg of one or more allergen proteins(s), e.g., between 1 ng and1.5 mg of one or more allergen protein(s), e.g., between 100 ng and 100μg of one or more allergen protein(s). In some embodiments, acomposition of the invention (e.g., a composition for inclusion in aflexible porous pouch or a dental hygiene product or chewing gum) can beformulated so that typical use provides (e.g., on a daily basis or overa period of time such as a week) between 0.01 and 100 times an amount ofallergen useful in a SLIT regimen, e.g., between 0.1 and 10 times, orbetween 0.5 and 2 times an amount useful in a SLIT regimen. For example,in some embodiments, a composition (e.g., dental hygiene product)comprising Cry j 1 and/or Cry j 2 can provide between 20 JAU and 20,000JAU on a weekly basis, e.g., between 200 JAU and 2000 JAU. In someembodiments, a composition contains 1, 2, 3, 4, 5, or more allergenprotein(s), wherein each allergen protein may independently be presentin any of the afore-mentioned quantities and/or the total amount ofallergen present may be any of the afore-mentioned quantities. One ofskill in the art would readily select an effective amount for inclusionin a composition or product of the invention. Homeopathic amounts ofallergen are also contemplated for use in inventive compositions (e.g.,dental hygiene product). For example, a 30c homeopathic dilution of anallergen extract or other allergen preparation can be used. As known inthe art, a 30c dilution represents 30 sequential 1 in 99 dilutions ofthe stock solution. In some embodiments, a 15c to 60c dilution is used.In some embodiments, the resulting solution theoretically contains nomolecules of allergen or of the original solution.

Methods for decreasing sensitivity to one or more allergens and reducingsymptoms of allergy in a subject are also provided. The method comprisesproviding sustained exposure to one or more allergens associated withthe allergy on an oral mucosal surface located substantially in avestibular region of the oral cavity of the subject. In certainembodiments the sustained exposure is provided by insertion of aflexible porous pouch capable of fixedly conforming to a vestibularmucosal surface of the oral cavity wherein the pouch contains acomposition according to the present invention, e.g., the nutraceuticalcomposition or pharmaceutical composition according to the invention. Itwill be appreciated that the buccal and/or gingival mucosa may also beexposed to allergen through use of inventive delivery forms. Furtherprovided are flexible porous pouches capable of conforming to the buccalmucosa wherein the pouch contains a composition according to the presentinvention.

In some aspects, methods of treating a subject suffering from or at riskof an allergic condition (e.g., allergy to one or more allergensdisclosed herein) are provided, wherein a method comprises prescribing,recommending, or suggesting use of an inventive composition or productto the subject. Optionally the method comprises determining that thesubject is allergic to an allergen, e.g., based on history and/or one ormore diagnostic tests. In some aspects, methods of treating a subjectsuffering from or at risk of an allergic condition are provided (e.g.,self-treatment methods), wherein a method comprises use of an inventivecomposition or product by the subject. A subject or may be aware ofparticular allergens that trigger an allergic reaction and selects aparticular composition or product based on such awareness.

In other specific embodiments sustained exposure is provided by ordinaryuse of a dental hygiene product such as toothpaste, dental cream,mouthwash or mouthspray. The invention provides suitable toothpaste,dental cream, mouthspray and mouthwash composition embodimentscomprising base ingredients of the dental hygiene product and furthercomprising one or more allergens. The one or more allergens may beprovided in suitably ground or refined natural form, or as processedinto an extract of the allergen or otherwise at least partially purifiedor isolated (e.g., separated from one or more substances with which itnaturally occurs or produced by man (e.g., using recombinant DNAtechnology)). The former specific embodiment contemplates that thenatural form is non-toxic and safe for inadvertent consumption. Allergenextracts processed specifically for safety and efficacy in all forms ofAIT are well known in the art and available from commercial vendors suchas GREER Laboratories (see, supra). Certain proprietary allergen blendsspecifically intended for nutraceutical compositions are particularlysuited to these embodiments. As a non-limiting illustrative example,proprietary plant blends intended as nutraceutical food additives areknown, including Pantescal® available from Bionap, Italy, which havedemonstrated immunosuppressive efficacy in individuals with certaingrass allergies, and are suitable for inclusion in this and otherembodiments of the instant invention.

Base formulations for dental hygiene products are well known to a personof skill in the art. One illustrative example of a base toothpastecomposition includes ingredients which comprise a combination of knownamounts of: vegetable glycerin; sorbitol; hydrated silica; purifiedwater; xylitol; carrageenan; sodium lauryl sulfate; titanium dioxide;propylparaben; methylparaben; sodium benzoate; and a flavoring agent.One of ordinary skill in the art will appreciate that toothpaste canencompass a paste, gel, or combination thereof. One of ordinary skill inthe art will also appreciate that a variety of ingredients may beincluded in an inventive toothpaste to serve, for example, asthickening/gelling agents, humectants, surfactants, flavoring orsweetening agents, polishing agents, etc. For example, thickening agentsinclude various cellulose derivatives (e.g., cellulose ethers such ascarboxymethyl cellulose), starch, gum (e.g., gum tragacanth, xanthangum), carrageenan, and silica-based thickening agents. Humectantsinclude, e.g., glycerin, glycols (e.g., propylene glycol, polypropyleneglycol, polyethylene glycol). A surfactant (which may or may not bepresent) may be anionic, nonionic, amphoteric, zwitterionic, cationic,or mixtures thereof. Exemplary anionic surfactants include, for example,water-soluble salts of alkyl sulfates having from 8 to 20 carbon atomsin the alkyl radical (e.g., sodium alkyl sulfate) and the water-solublesalts of sulfonated monoglycerides of fatty acids having from 8 to 20carbon atoms. Exemplary nonionic surfactants include triblock copolymerscomposed of polyoxypropylene and polyoxyethylene, for example,poloxamers (sold under the trade name Pluronics, for example).Sweetening agents include, e.g., saccharin, xylitol, sorbitol,aspartame. Polishing agents include a variety of different abrasivematerials and may be silica-based or non-silica based (e.g., calcium oraluminum-based, e.g., calcium carbonate, alumina, aluminum hydroxide).Flavoring agents include a variety of plant-derived oils, for example.In some embodiments, water, thickening/gelling agent, polishing agent,humectant, surfactant together amount to between 70% and 99.99% byweight of the composition. In some embodiments, water,thickening/gelling agent, polishing agent, humectant, and/or surfactantis present at between 5% and 70% by weight, with the total of theseingredients together amounting to between 70% and at least 99.99% byweight of the composition. In some embodiments, a polishing agent ispresent in an amount between 5% and 50% by weight, based on the totalweight of the composition. In some embodiments, a surfactant is presentin amount between 0.5% and 15% by weight, e.g., 1.0% to 10% by weight,based on the total weight of the composition.

In some embodiments, a dental hygiene product, e.g., toothpaste,comprises an anti-caries agent such as a soluble fluoride source capableof providing free fluoride ions, while in some embodiments a fluoridesource is not included. Soluble fluoride ion sources include sodiumfluoride, stannous fluoride, and sodium monofluorophosphate, forexample. See, e.g., U.S. Pat. Nos. 2,946,725 and 3,678,154. In someembodiments, fluoride ion source is provided in an amount sufficient toprovide between 50 and 3500 ppm, e.g., between 500 and 1500 ppm,fluoride ions. A non-fluoride anti-caries agent (e.g., anti-bacterialagent) may be included or not. Other ingredients that may be present insome embodiments or specifically excluded include a bicarbonate ionsource, a pyrophosphate ion source (e.g., pyrophosphate salt), bufferingagent(s), coloring or opacifying agent(s) (e.g., titanium dioxide), astabilizer, and preservative(s) (e.g., parabens). Representativeexamples of various dental hygiene product formulations, ingredients,and methods of preparation are found, for example, in PCT/US1997/021157(WO/1998/023250) FLAVOR SYSTEMS FOR ORAL CARE PRODUCTS and in Weinert,W., “Oral Hygiene Products”, Ullmann's Encyclopedia of IndustrialChemistry, Wiley-VCH, Weinheim, 2000. One of ordinary skill in the artwould readily select ingredient(s) and formulation approaches that arecompatible with each other (e.g., do not significantly reduce thecapacity of an ingredient to produce the intended effect). One ofordinary skill in the art will appreciate that some ingredients mayserve multiple functions (e.g., sorbitol may serve as a sweetener andhumectant) and that various categories of ingredient may be omitted orother categories may be included. The invention contemplates “natural”toothpastes that are essentially devoid of artificially synthesizedcompounds that do not occur in nature.

The toothpaste or dental cream of this invention is typically preparedby conventional methods of making toothpastes and/or dental creams ordental gels (with the proviso that an allergen is incorporated into thecomposition). More specifically, for example, in some embodiments thegelling agent such as a cellulose gum is dispersed in glycerine, towhich is added an aqueous solution containing the sweetening agent suchas xylitol, followed by the addition of sorbitol and mixing for a periodof about 20 minutes to hydrate the gum, mixing the gum mixture with thepolishing agent in a mixer under a vacuum of 28-30 inches of pressure.Lastly, the flavor, the surfactant and processed allergen are added tothe vacuum mixer, mixed for a period of about 15 minutes, and the finalmixture is placed in a tube or other container suitable for dispensingthe toothpaste, such as a vertical dispenser equipped with a pump. Ofcourse ingredients can be combined in any appropriate order compatiblewith the nature of the ingredients. The tube, pump, or other dispensingcontainer may be marked to indicate that its contents include anallergen and/or to depict one or more allegen(s) or allergen source(s)and/or may feature a brand name, logo, etc. The container may be labeledwith information regarding flavor, ingredients, etc. Similarconsiderations apply with regard to packaging, labeling of otherinventive delivery forms, e.g., chewing gum, dental pick or floss,porous pouch, etc.

An ordinary use involves, for example, approximately 2 gm of toothpastehaving approximately 1-10% by weight processed allergen (or othersuitable amount of allergen in processed or natural form, as discussedabove). In some embodiments a “pea-sized” quantity of toothpaste (e.g.,0.2 gm-0.5 gm, e.g., about 0.3 gm) may be suitable for a child betweenthe ages of 2 and 6 years. In some embodiments, a toothpaste dispenserthat dispenses a predetermined quantity (e.g., a predetermined volume)of toothpaste upon activation is used. Activation may be achieved, e.g.,by pushing a button, depressing a lever, activating a motion sensor, orother approaches as known in the art.

In certain embodiments the toothpaste may be divided into portions, afirst portion comprising ingredients relating to oral cleanliness andhealth, and a second portion comprising ingredients relating to thedevelopment of allergic tolerance. Production of multi-portionedtoothpastes wherein, for example, the portions are coaxially aligned indelivery form, are well-known in the art. This product delivery form istypically employed where composition requirements vary as between thebase ingredients and desired additive ingredients, or where it isdesired that the ingredients of one portion remain in the oral cavityfor a longer period of time. For example, whereas a portion comprisingbase ingredients may be hydrophilic and substantially removed by rinsingof the oral cavity with water, a second portion comprising allergenicextract may be hydrophobic and may include filming properties. In someembodiments, an inventive toothpaste is multi-striped, wherein thedifferent stripes differ in composition. For example, one or morestripes may contain allergen while others do not. In some embodiments, amulti-cavity dispensing container that permits simultaneous coextrusionof two or more flowable materials, e.g., in a predetermined proportion,is used. See, e.g., U.S. Pat. No. 5,020,694. In some embodiments, thetoothpaste has a multilayer composition. In producing such embodiments,desired toothpaste layers can be fed into the dispensing container(e.g., tube) in parallel streams to form a multilayered appearance. Thedentifrice layers will be extruded in the desired multilayerconfiguration when dispensed from the tube.

A person of ordinary skill in the art of dental hygiene productformulation will immediately recognize how to match a compositioningredient characteristic to a formulation parameter to achieve desiredeffect with respect to exposure time in the oral cavity.

Unique advantages afforded by this embodiment include very highpotential for full compliance by the consumer. In conventional andordinary usage, the typical American already purchases dental hygieneproducts for use several times per day. Over 40 percent of Americansreport that brushing teeth just prior to bedtime is a ritual or habitualactivity. Hence, the enhanced exposure is accomplished as part of analready existing daily routine. In some embodiments, a daily regimenincludes at least one exposure of the inventive dental hygiene productto the oral cavity (e.g., 1, 2, 3, 4, or 5 exposures) for periodsranging from 15 seconds to 10 minutes. A contemplated suitable useregimen includes exposure of the inventive dental hygiene product to theoral cavity 2-4 times per day for periods ranging from about 30 to about90 seconds. Regimens involving less frequent use are also contemplated,e.g., 1, 2, or 3 times weekly, alternate days, etc.

The toothpaste can be applied using a typical toothbrush, e.g., a manualtoothbrush comprising a head with bristles, a handle for gripping thetoothbrush, and a neck portion connecting the head and the handle, or anelectric toothbrush. An electric toothbrush may additionally comprise amotor and a power source (e.g., a battery-operated, optionallyrechargeable) in electrical communication with the motor. In someembodiments, a toothbrush comprises a timer that produces a signal(e.g., a sound or light) after a brushing session has continued for atleast a predetermined period of time. For example, the timer may producea signal after 30, 60, 90, 120, 150, 180, 210, or 240 seconds, invarious embodiments. The period of time may be selectable, e.g., by theuser. Of course a timer that is separate from the toothbrush could beused, if desired. In some embodiments, the toothbrush briefly interruptspower at predetermined intervals or after a predetermined interval,e.g., any of the afore-mentioned time periods, thereby prompting theuser to begin brushing a different area of the mouth.

In some embodiments, an electric toothbrush comprises one or moremicroprocessor(s), data storage element(s), and/or display(s). Themicroprocessor(s), data storage element(s) (“memory”), and display(s)can be typical components as commonly found in consumer electronicdevices. The display can comprise, for example, a LCD or LED screen. Thememory can store information regarding, for example, the duration,frequency, and/or other characteristic(s) of tooth brushing sessions.Such data can be displayed on the toothbrush display or on a separatedisplay. A microprocessor, data storage element, and/or display may, forexample, be located within the toothbrush, within the base unit of anelectric toothbrush, or in a completely separate unit. In someembodiments, a display prompts a user to continue brushing or ceasebrushing and/or provides other messages to the user. PCT/US2007/023677(WO/2008/060482—PERSONAL CARE PRODUCTS AND METHODS) discloses anillustrative oral care system comprising an electric toothbrush and adisplay in data communication with the electric toothbrush.

In some embodiments, a tooth brushing system, e.g., a toothbrush, servesas a dispenser for an inventive dental hygiene product and/or method.Tooth brushing systems, e.g., toothbrushes, suitable for dispensingtoothpaste and/or other oral care products are known in the art. Thetoothbrush can comprise one or more reservoir(s) from which toothpasteand/or or other material(s) is/are dispensed prior to and/or duringbrushing, or a brush may be removable from a base that contains one ormore reservoir(s). Dispensing can, for example, occur prior to a toothbrushing session and/or intermittently or continuously during at leastpart of a tooth brushing session. In some embodiments, a metered dosingpump, which may be included in the body of the toothbrush, controls theflow of toothpaste from the reservoir and through a conduit thatdelivers the toothpaste to the toothbrush head, e.g., to or adjacent tothe bristles of the toothbrush head. Operation of the metered dosingpump dispenses a measured and controlled amount of toothpaste. In someembodiments, a first reservoir comprises toothpaste, and a secondreservoir delivers additional material (e.g., an allergen preparation)to the flow of toothpaste. In some embodiments, an allergen (optionallytogether with one or more other components) is delivered at least inpart after the toothpaste has been dispensed. The allergen preparationmay, for example, be delivered to the bristles or through the toothbrushhead. In some embodiments, conventional toothpaste is delivered from afirst reservoir, and an allergen is delivered from a second reservoir.In some embodiments, a toothbrush is loadable with cartridge(s) thatcontain an allergen. Such allergen-containing cartridges suitable forinserting into a toothbrush or using to load a toothbrush with anallergen preparation are an aspect of the invention. Delivery oftoothpaste and/or allergen preparation may be regulated by amicroprocessor, which is optionally programmable to deliver a selectedamount of toothpaste and/or allergen preparation. Illustrative examplesof tooth brushing systems capable of serving as a dispensing system aredisclosed in PCT/US2001/043442 (WO/2002/041802—APPARATUS, METHOD ANDPRODUCT FOR TREATING TEETH) and PCT/US2008/054695(WO/2008/103892—TOOTHBRUSH WITH INTEGRATED TOOTHPASTE DELIVERY. Atoothpaste packaged as a dual chambered container is disclosed inSoparkar P., et al., J Clin Dent. 15(2):46-51, 2004.

In some aspects, the invention provides a dental floss or dental pickthat delivers a composition comprising an allergen and/or is used topromote contact of a composition of the invention with the oral mucosa(e.g., gingival and/or vestibular mucosa). In some embodiments, a dentalpick comprises multiple flexible bristles (e.g., rubber bristles) that,for example, slide between teeth to remove plaque and food particles,while promoting circulation by massaging and stimulating the gingiva.For example, the dental pick may have about 50-100 bristles, e.g., about70-80 bristles. In some embodiments, a user applies a dental hygieneproduct of the invention (e.g., toothpaste, oral gel) to the oral mucosa(e.g., gingviva and/or vestibulum) and/or to the bristles of the dentalpick. Use of the dental pick distributes the composition and promotescontact with the oral mucosa. An exemplary dental pick is the GUM®Soft-Pick® (Sunstar Americas, Inc.).

In some embodiments, a dental floss comprises bundle of thin filaments(e.g., nylon filaments) or fibers, or a plastic (e.g., Teflon orpolyethylene) ribbon of use to remove food and dental plaque from teeth.In some embodiments a dental floss may be flavored or unflavored, andcoated or uncoated (e.g., waxed or unwaxed). Dental floss may beprovided in a dispenser or dental floss pick, for example. In someembodiments, a dental floss comprises an allergen impregnated into acoating layer (e.g., a wax) or distributed as an outer layer of acoating layer. In some embodiments, an allergen is encapsulated, e.g.,microcapsules or nanocapsules. In some embodiments, encapsulation helpsin stabilizing and/or preserving the potency of allergen, e.g., duringmanufacture and/or storage of the dental floss. In some embodiments,allergen desorbs or is otherwise released from the coating and/ormicrocapsules as the dental floss is used. Suitable materials forencapsulating active agents are known in the art and include a widevariety of different synthetic and naturally occurring polymers. In someembodiments, release of allergen is promoted by mechanical action offlossing between teeth and/or by contact with water and/or saliva in themouth, e.g., by enzymatic action, e.g., of enzymes in saliva.

In some embodiments, a dental pick or dental floss is packaged orotherwise sold or marketed together with an inventive dental hygieneproduct (e.g., a toothpaste, gel, etc.). Such packages are within thescope of the invention.

Another specific embodiment directed to providing a compositionaccording to the invention in very convenient and familiar delivery modeinclude application by use of an oral gel. This embodiment isparticularly suitable for night time exposure as salivation decreasedsubstantially at night, and the gel does not present the choking hazardinherent to solid delivery forms. Hence, in a very specific embodimentthe gel is applied to the vestibular mucosal surface just prior tosleeping. In some embodiments, the gel is applied at least in part tothe buccal mucosa just prior to sleeping. In other specific embodiments,the gel is provided in the form of a gel plug. The gel plug is designedto fit in any of several nooks of the vestibular region.

In a further specific embodiment, the sustained exposure is provided byan oral strip comprising the one or more allergens, wherein the oralstrip comprises a hydrophilic polymer that dissolves with exposure tosaliva. According to this embodiment, the oral strip may be applied tothe vestibular mucosal surface just prior to sleeping or until it fullydissolves. Guidance for production of suitable oral strips in accordancewith the instant invention is provided, for example, in R. P. Dixit etal. ‘Oral Strip Technology: Overview and Future Potential’ J. ControlledRelease 139 (2009) 94-107, the disclosure of which is incorporatedherein by this reference. Dixit teaches suitable formulation parametersand exemplifies additives which may considered including sweetening,flavoring, coloring, stabilizing and thickening agents and salivastimulating agents. Manufacturing concerns including strip-formingpolymers, thickness, industry tests, tensile strength, elongation, tearand fold parameters, dissolution and disintegration control factors areall addressed therein.

The invention provides chewing gums comprising one or more allergens. Ingeneral, a chewing gum of the invention may be produced usingconventional ingredients and methods for manufacturing chewing gums,with appropriate modifications to incorporate one or more allergens. Oneof ordinary skill in the art is aware of appropriate ingredients andformulation methods for chewing gums. For example, a chewing gum maycontain a gum base comprising chicle, a natural latex product, othernatural gums, or a synthetic rubber (e.g., polyisobutylene). Typicallychewing gum contains one or more flavoring agents, to impart a flavorsuch as mint, wintergreen, cinnamon, etc., and may contain sweeteningagent(s). Ingredients can include one or more humectants, softeners,coloring agents, stabilizers, preservative, etc. A chewing gum may havefilm-forming characteristics for blowing bubbles. Chewing gum can bemanufactured in a variety of shapes such as sticks, ribbons, balls, etc.In some embodiments, a chewing gum is coated or glazed, e.g., with asuitable wax. In some embodiments, a gum is a center-filled gum, e.g., apellet or ball gum formed around a soft or liquid center. In someembodiments, an allergen is provided in such center. Optionally, thecenter is surrounded at least in part by a barrier layer. Suitablebarrier layers may comprise, e.g., a wax. See, e.g., US patentapplication publication 20060263476 (CENTER-FILLED CHEWING GUM WITHBARRIER LAYER). In general, the amount of allergen to be incorporatedinto a stick of chewing gum can be as described above for pouch ordental hygiene product. In some embodiments, a typical use entailschewing 1-5 units (sticks, balls, etc.) of chewing gum daily. Regimensinvolving less frequent use are also contemplated, e.g., 1, 2, or 3times weekly, alternate days, etc. In some embodiments, a typical useentails chewing 1-10 units of chewing gum weekly.

In some aspects, the invention provides a method of making a dentalhygiene composition (e.g., toothpaste, mouthwash, dental cream), themethod comprising: a) providing base ingredients for said dental hygienecomposition; and b) combining at least one allergen with said baseingredients. In some embodiments, the method further comprises (c)placing said dental hygiene composition in a container suitable fordispensing the composition. In some embodiments, the method comprisesproviding the at least one allergen in a form that has been quantifiedwith regard to allergen content. The allergen may, for example, havebeen quantified based on a reference standard. In some embodiments, themethod comprises providing the at least one allergen in an amountappropriate to result in a predetermined minimum and/or maximum amountof allergenic protein and/or predetermined minimum and/or maximum numberof allergen units, e.g., on a volume or weight basis of the dentalhygiene composition. In some embodiments, a method comprises testing thedental hygiene composition, e.g., to determine the amount of allergen inthe composition, e.g., to confirm that the composition comprises atleast a predetermined minimum amount of allergen and/or that the amountof allergen does not exceed a predetermined maximum amount, e.g., on avolume or weight basis of the dental hygiene composition. In someaspects, the invention provides dental hygiene composition and methodsof making thereof, wherein the dental hygiene composition has batch tobatch consistency with regard to allergen content.

In some aspects, similar methods are provided for making a chewing gumof the invention. In some embodiments, an allergen comprises a protein,and the allergen is substantially the only protein ingredient includedin the dental hygiene composition or chewing gum. In some embodiments,an allergen comprises a protein, and the allergen is structurallydistinct from any ingredients that may be conventionally used in adental hygiene composition or chewing gum.

In some embodiments of any aspect of the invention (e.g., dental hygieneproduct such as toothpaste; dental pick or dental floss composition forinclusion in porous pouch, chewing gum), an allergen can be associatedwith microparticles. For example, the allergen can be encapsulated bysuch particles and/or microparticles can be impregnated or coated orotherwise physically associated with allergen. “Microparticle” as usedherein, encompasses any microscopic particles used to protect and/ordeliver agents in areas such as pharmaceuticals, nutraceuticals,cosmeceuticals, cosmetics, food technology, and the like. Such particlesmay be referred to in the art as microcapsules, microspheres,nanospheres, nanoparticles, nanocapsules, liposomes, and the like.Methods of making and employing such delivery systems are well known inthe art. Examples are described, e.g., in references such as: Lakkis, JM (ed.) Encapsulation and controlled release technologies in foodsystems (Wiley-Blackwell, 2007); Nedovic, V. and Zuidam, N J (eds.)(Springer, 2009); Cohen, S. and Bernstein, H. Microparticulate systemsfor the delivery of proteins and vaccines (CRC Press, 1996); Jones, D.,Pharmaceutical Applications of Polymers for Drug Delivery (ChemTecPublishing, 2004). Benita, S. (ed.) Microencapsulation: methods andindustrial applications (Informa Healthcare; 2^(nd) ed., 2005), all ofwhich are incorporated by reference. In some aspects, known approachesused in substances expected to contact the oral cavity (e.g., foods) areused in the present invention. A number of polymeric delivery vehiclesfor providing sustained release are known in the art. One of ordinaryskill would select appropriate polymers for use in various embodimentsof the invention. In some embodiments, a biocompatible polymer, whichmay be biodegradable, may be used. The polymers may be homopolymers,copolymers (including block copolymers), straight, branched-chain, orcross-linked. Natural or synthetic polymers can be used in variousembodiments of the invention. Polymers include, but are not limited to,poly-lactic acid (PLA), poly-glycolic acid (PGA),poly-lactide-co-glycolide (PLGA), poly(phosphazine), poly (phosphateester), polycaprolactones, polyanhydrides, ethylene vinyl acetate,polyorthoesters, polyethers, and poly (beta amino esters). Otherpolymers include polyamides, polyalkylenes, polyalkylene glycols,polyalkylene oxides, polyalkylene terepthalates, polyvinyl alcohols,polyvinyl ethers, polyvinyl esters, poly-vinyl halides,polyvinylpyrrolidone, polyglycolides, polysiloxanes, polyurethanes andco-polymers thereof, poly(methyl methacrylate), poly(ethylmethacrylate), poly(butylmethacrylate), poly(isobutyl methacrylate),poly(hexylmethacrylate), poly(isodecyl methacrylate), poly(laurylmethacrylate), poly(phenyl methacrylate), poly(methyl acrylate),poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecylacrylate), polyethylene, polypropylene, poly(ethylene glycol),poly(ethylene oxide), poly(ethylene terephthalate), poly(vinylalcohols), polyvinyl acetate, poly vinyl chloride, polystyrene,polyvinylpyrrolidone, poly(butyric acid), poly(valeric acid), andpoly(lactide-cocaprolactone). Peptides, polypeptides, proteins such ascollagen or albumin, polysaccharides such as sucrose, chitosan, dextran,alginate, hyaluronic acid (or derivatives or combinations of any ofthese), dendrimers (e.g., PAMAM dendrimers), dextrins, cyclodextrins maybe used in various embodiments. Methods for preparation of suchformulations will be apparent to those skilled in the art. Liposomes orother lipid-containing particles can be used in some embodiments.Exemplary polymers include cellulose derivatives such as, alkylcellulose, hydroxyalkyl celluloses, cellulose ethers, cellulose esters,nitro celluloses, polymers of acrylic and methacrylic esters, methylcellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxy-propylmethyl cellulose, hydroxybutyl methyl cellulose, cellulose acetate,cellulose propionate, cellulose acetate butyrate, cellulose acetatephthalate, carboxymethylcellulose, carboxylethyl cellulose, cellulosetriacetate, cellulose sulphate sodium salt, polycarbamates or polyureas,Chemical derivatives of the afore-mentioned polymers, e.g.,substitutions, additions of chemical groups, for example, alkyl,alkylene, hydroxylations, oxidations, and other modifications routinelymade by those skilled in the art can be used.

In some embodiments of the invention, a composition for use in adelivery form of the invention (e.g., a pouch or a dental hygieneproduct or chewing gum) contains one or more allergen(s) of relevance toa particular geographic area. For example, the allergen(s) can beallergen(s) of plants (e.g., pollen allergens) that grow commonly insuch area, and/or allergen(s) of fungi, insects, mites, etc., which arecommonly found in such area. In some embodiments, a composition for usein a delivery form of the invention (e.g., a pouch or a dental hygieneproduct) contains one or more allergen(s) that are significant causes ofallergy in a geographical area. In some embodiments, an allergen is asignificant cause of allergy if it is among the 5 most commonlydiagnosed allergy-causing agents in a particular category (e.g.,airborne, ingested, skin contact) in a geographical area. Diagnosis maybe based on, e.g., skin prick testing or other methods accepted in theart. A geographical area can be a continent, e.g., North America, SouthAmerica, Europe, Asia, Africa, Australia; one or more countr(ies),regions, or jurisdictions (e.g., US, Canada, Mexico, Argentina, Brazil,Chile, Venezuela, European Union, Belgium, Denmark, France, Germany,Italy, Netherlands, Norway, Poland, Spain, Sweden, Switzerland, UnitedKingdom, Turkey, Russia, Eurasia, Israel, Japan, China, Korea, India,Pakistan, Philippines, Singapore, Vietnam, Thailand, Indonesia, Egypt,South Africa, ARIPO member state(s), Australia, etc.) or portion(s)thereof (e.g., one or more states or provinces). A geographical regioncan be defined based at least in part on climate or other naturalfeatures. In some embodiments a geographical area is at least 10,000 km²in area. In some embodiments, a dental hygiene product of the inventionis formulated to have one or more characteristic(s) (e.g., flavor,color, texture, etc.) that closely match those of a dental hygieneproduct that is familiar to at least some people living in a particulargeographic area that corresponds to the allergen(s) in the inventivedental hygiene product, e.g., a dental hygiene product that is marketedin such area. In some aspects, the invention provides methods of makinga dental hygiene composition comprising a) providing base ingredientsfor said dental hygiene composition; and b) combining at least oneallergen with said base ingredients, wherein the at least one allergenis relevant to a selected geographical area. In some embodiments thebase ingredient(s) and/or amount(s) thereof are selected such thatcharacteristic(s) of the resulting composition are familiar to at leastsome people living in such geographic area.

In some embodiments of the various aspects of the invention, a subjectis an individual who has a history of allergy symptoms and/or who hasreceived a diagnosis of allergy. In some embodiments, a subject is atrisk of developing an allergy. In some embodiments a subject who has afamily history of allergy is at risk of developing an allergy. In someembodiments, a subject has a positive skin prick test or skin patch testto one or more allergen(s). In some embodiments, a subject hasdetectable serum IgE (as assessed, e.g., using RAST) against one or moreallergen(s). In some embodiments, a subject who has detectable serumIgE, e.g., abnormally high levels of serum IgE (as assessed, e.g., usingRAST) against an allergen is at risk of developing an allergy to thatallergen (or to other allergens that share cross-reactive epitopes.) Asknown in the art RAST (radioallergosorbent test) is a radioimmunoassaytest used to detect specific IgE antibodies to suspected or knownallergens. Since IgE is the antibody associated with Type I allergicresponse, if a subject exhibits a significant level of IgE directedagainst an allergen, the test may indicate that the subject is allergicto such allergen (or to allergens that share epitopes with the allergen)or is at risk of developing an allergy to the allergen (or to allergensthat share epitopes with the allergen). Other methods of detectingallergen-specific IgE are known in the art.

As known in the art, many allergens are immunologically cross-reactivewith other allergens, e.g., allergens that share at least some similaror identical epitopes (e.g., IgE antibodies that bind to a firstallergen will also bind to allergens that share at least some similar oridentical epitopes). Thus an individual who is allergic to an allergenfrom a first species will often be allergic to other allergens, e.g.,allergens from related species. In some embodiments, a first allergenmay be used to desensitize a subject to a second allergen. The first andsecond allergens may be derived from different sources. For example, inthe case of allergens of plant, animal, or fungal origin, the first andsecond allergens may be derived from different species within a genus,or different genera within a subfamily or family, or differentsubfamilies within a family. For example, in some embodiments, theinvention contemplates use of a pollen allergen from one or more grassesor trees of a first subfamily to desensitize individuals to pollen ofone or more grasses or trees of a second subfamily. In some embodiments,the invention contemplates use of pollen allergen from trees of thesubfamily Cupressoideae (e.g., junipers) to desensitize individuals topollens of the subfamily Taxodioidea (e.g., Japanese cedar), or viceversa. For example, Japanese cedar pollen allergens can beimmunologically cross reactive with allergens of Cupressus sempervirens,Juniperus ashei, Cupressus arizonica, Cupressus macrocarpa, Juniperusvirginiana, Juniperus communis, Thuya orientalis, and/or Chamaecyparisobtusa.

Personalized methods for decreasing a subject's allergic response to anenvironmental allergen are also provided. In certain embodiments anallergen associated with a subject's allergic response is identified anda commercially available extract of the allergen is procured. Theextract is mixed with, for example, an organic non-toxic fillermaterial. A flexible porous pouch according to the invention is filledwith the mixture, wherein the filled pouch is suitable for insertion bythe subject into an oral cavity of the subject and capable of fixedlyconforming to a vestibular mucosal surface of the oral cavity, andinserting the pouch so that it fixedly conforms to the vestibularmucosal surface. It is contemplated that the identified allergen derivesfrom the subject's environment and is suspected of causing undesirableclinical symptoms in the subject. In very specific embodiments thesubject may be able to avoid the prospect of having to eliminate apleasurable source of the offending allergen, for example a pet, bytailoring vestibular immunotherapy with respect to the particular pet.

Any of the aspects, embodiments, and features of the invention can befreely combined, and such combinations are within the scope of theinvention. For example, in some aspects, the invention contemplates useof multiple different inventive delivery forms by a subject. Forexample, a subject may use an inventive pouch containing an allergenduring an induction phase and a dental hygiene product, e.g., atoothpaste, for maintenance. Inventive delivery forms can also be usedtogether with SLIT or other forms of immunotherapy. For example, asubject may initiate desensitization with SLIT and switch to use of aninventive delivery form for maintenance.

In general, a subject could start on a regimen of using an inventivepouch, dental hygiene product, or chewing gum at any time. In someembodiments, it is contemplated that a subject starts on a regimen ofusing an inventive pouch, dental hygiene product, or chewing gum atleast 7 days prior to an anticipated exposure to an allergen. Forexample, a subject may commence using an inventive pouch, dental hygieneproduct, or chewing gum at least 1-4 weeks, 4-8 weeks, 8-16 weeks, 16-24weeks, 24-36 weeks, or 36-52 weeks prior to an anticipated exposure,etc. “Rush” regimens and “ultra-rush” regimens are also contemplated. Ina “rush regimen,” a subject is exposed to successively increasing amountof allergen spaced over a relatively short period of time such as about8 hours to 7 days. In an “ultra-rush” regimen, a subject is exposed tosuccessively increasing amount of allergen separated by short timeintervals, such as 30 minutes, over a period of about 2-8 hours. Ananticipated exposure may be, for example, the typical beginning ofpollen season, a planned trip to a location where the allergen occurs inthe environment, acquisition of a pet, etc. The subject may continueusing inventive pouches or dental hygiene products during the period ofexposure to the allergen. In some embodiments, a subject uses aninventive product prior to and during multiple seasonal exposures to anallergen, e.g., a pollen. In some embodiments, a pre-coseasonal protocolis used, which starts every year approximately 4 months before thepollen season and is continued throughout the season (approximately 1 to2 months depending on the country and regions) and then is stopped atthe end of the season. This will be repeated each year approximately 4months before the pollen season in accordance with the same protocol. Insome embodiments, a perennial protocol is used, which can be started atany time (e.g., at least 4 months before the pollen season) andtypically involves regular use (e.g., daily, weekly, etc.) continuouslythroughout the course of use. In some embodiments, the overall durationof use is 3 to 5 years or 3 to 5 consecutive seasons in the case of thepre-coseasonal protocol. In some embodiments, it is contemplated thatthe subject continues to use inventive pouches or dental hygieneproducts indefinitely. In some embodiments, a protocol includes aninduction phase that involves escalating the amount of deliveredallergen over a period of time to reach a maintenance dose level.

Any of the inventive delivery forms or compositions may be packaged orotherwise provided together with instructions, e.g., relating tosuitable use regimens, amounts, etc.

In some embodiments, an inventive pharmaceutical composition (e.g., acomposition for inclusion in a pouch, or the pouch containing thecomposition, or a dental hygiene product or chewing gum) is packagedtogether with a label approved by a government agency responsible forregulating pharmaceutical agents, e.g., the U.S. Food & DrugAdministration, or comparable agency in a different jurisdiction.

The present invention also provides kits. A kit comprising (a) aplurality of flexible porous pouches suitable for insertion into an oralcavity of a subject and capable of fixedly conforming to a vestibularmucosal surface of the oral cavity, the pouches comprising a compositioncomprising at least one allergen and (b) instructions comprising adosing schedule effective for desensitizing the subject to the at leastone allergen thereby reducing the severity of at the clinical symptoms.

In some embodiments, the invention provides a kit comprising: aplurality of flexible porous pouches as described herein; a plurality ofindividually contained extracts of a set of common environmentalallergens; an amount of organic non-toxic filler material; andinstructions for mixing extract with the filler material and for fillingthe flexible porous pouch.

In some embodiments, the invention provides a kit as described herein,wherein the composition of the set of common environmental allergens isbased on the environment of the subject.

If desired, the effect of the inventive allergen immunotherapy productswith regard to desensitizing a subject can be assessed using methodsknown in the art for assessing symptoms and/or signs of allergy. In someembodiments, an effective amount is an amount that decreases(ameliorates, reduces, etc.) one or more symptoms or manifestations ofallergy and/or decreases likelihood of developing one or more symptomsor manifestations of allergy (or associated conditions such as asthma,atopic dermatitis) upon exposure to an allergen, e.g., as compared withthe level of such symptom(s) or manifestation(s) expected or previouslyexperienced by a subject in the absence of using an inventivecomposition or product of the invention. For example, subject becomessignificantly less sensitive to the allergen than previously the case,or than would be expected based, for example, on the subject's history.In some embodiments, a reduction in symptoms is clinically significant,e.g., as assessed by a medical practitioner familiar with the allergiccondition. One of ordinary skill in the art would be aware of symptomsand manifestations of allergic conditions. Typical allergy symptoms(e.g., to airborne allergens) include, for example, sneezing,rhinorrhea, nasal congestion, nasal and ocular pruritus (itching), andtearing. Further details regarding symptoms, diagnosis, etc., aredescribed in standard textbooks such as Adkinson, N F, et al.,Middleton's Allergy: Principles and Practice, 7^(th) edition (Mosby,2008) Symptoms can be scored, optionally using any of a variety ofinstruments known in the art for assessing allergy severity and/or forassessing efficacy of agents or procedures for alleviating allergicmanifestations. For example, in some embodiments, theRhinoconjunctivitis Total Symptom Score (RTSS), a sum of 6 individualsymptom scores for sneezing, runny nose, itchy nose, nasal congestion,watery eyes, and itchy eyes is used. See, e.g., Wahn U, et al., JAllergy Clin Immunol., 123(1):160-166, 2009, incorporated by reference.In some embodiments, the Japanese Allergic Rhinitis Quality of Life(QOL) Standard Questionnaire No. 1 (JRQLQ No. 1) (Okubo, supra) may beused. One of ordinary skill in the art will appreciate that manydifferent scoring systems suitable assessing allergy symptoms could beemployed. Use of rescue medications (e.g., antihistamines) can bemonitored, wherein a reduction in the requirement for rescue medicationsis indicative of a beneficial effect resulting from use of an inventiveproduct. Other methods of assessing efficacy include performing anantigen challenge test, e.g., by skin exposure (e.g., skin prick),respiratory exposure, ingestion, etc., and assessing response thereto ascompared, for example, with response prior to embarking on a regimen ofusing an inventive pouch or dental hygiene product. Appropriatestatistical test(s) (e.g., t-test, Chi-square test, ANOVA, etc.) knownto those of ordinary skill in the art can be used to demonstrate astatistically significant benefit, e.g., reduction in one or moresymptoms, reduction in total score on a standardized instrument,reduction in use of rescue medications, etc., achieved by using aninventive product. In some embodiments, statistical significance refersto a p value of <0.05. In some embodiments, statistical significancerefers to a p value of <0.01.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. The scope of the presentinvention is not intended to be limited to the above Description, butrather is as set forth in the appended claims. It will be appreciatedthat the invention is in no way dependent upon particular resultsachieved in any specific example or with any specific embodiment.Articles such as “a,” “an” and “the” may mean one or more than oneunless indicated to the contrary or otherwise evident from the context.Claims or descriptions that include “or” between one or more members ofa group are considered satisfied if one, more than one, or all of thegroup members are present in, employed in, or otherwise relevant to agiven product or process unless indicated to the contrary or otherwiseevident from the context. The invention includes embodiments in whichexactly one member of the group is present in, employed in, or otherwiserelevant to a given product or process. The invention also includesembodiments in which more than one, or all of the group members arepresent in, employed in, or otherwise relevant to a given product orprocess. Furthermore, it is to be understood that the inventionencompasses all variations, combinations, and permutations in which oneor more limitations, elements, clauses, descriptive terms, etc., fromone or more of the listed claims or from the description above isintroduced into another claim. For example, any claim that is dependenton another claim can be modified to include one or more elements,limitations, clauses, or descriptive terms, found in any other claimthat is dependent on the same base claim. Furthermore, where the claimsrecite a composition, it is to be understood that methods of using thecomposition for any of the purposes disclosed herein are included withinthe scope of the invention, and methods of making the compositionaccording to any of the methods of making disclosed herein are includedwithin the scope of the invention, unless otherwise indicated or unlessit would be evident to one of ordinary skill in the art that acontradiction or inconsistency would arise. Methods can include a stepof providing a subject having an allergy or being at risk of developingan allergy or allergic reaction, a step of diagnosing a subject ashaving an allergy or being at risk of developing an allergy or allergicreaction, and/or a step of selecting a subject for which an inventiveproduct or method would be suitable.

Where elements are presented as lists, it is to be understood that eachsubgroup of the elements is also disclosed, and any element(s) can beremoved from the group. For purposes of conciseness only some of theseembodiments have been specifically recited herein, but the inventionincludes all such embodiments. It should also be understood that, ingeneral, where the invention, or aspects of the invention, is/arereferred to as comprising particular elements, features, etc., certainembodiments of the invention or aspects of the invention consist, orconsist essentially of, such elements, features, etc.

Where ranges are given, endpoints are included. Furthermore, it is to beunderstood that unless otherwise indicated or otherwise evident from thecontext and understanding of one of ordinary skill in the art, valuesthat are expressed as ranges can assume any specific value or subrangewithin the stated ranges in different embodiments of the invention, tothe tenth of the unit of the lower limit of the range, unless thecontext clearly dictates otherwise. Any particular embodiment, aspect,element, feature, etc., of the present invention, or any combinationthereof, may be explicitly excluded from any one or more claims whetheror not such exclusion is expressly recited herein. For example, anyallergen or ingredient, etc., can be explicitly excluded. Applicantsreserve the right to proviso out of the claims any specific allergen,allergen category, ingredient, ingredient category, or combinationthereof, whether or not such allergen, ingredient, category, orcombination thereof, is recited herein. To the extent, if any, that adental hygiene product, chewing gum product, or a composition for use ina pouch that is known or described in the prior art may include anallergen, the instant invention may be distinguished from such prior artproduct or composition in, for example, any one or more of the followingways: (i) the product or composition of the invention comprises one ormore allergen(s) not present in the prior art product or composition;(ii) the product or composition of the invention comprises a differentamount of allergen, or a different form of the allergen, than present inthe prior art composition; (iii) the product or composition of theinvention explicitly excludes the allergen(s) present in the prior artproduct or composition; (iv) the composition of the invention comprisesat least one ingredient not present in the prior art product compositionor present in a different amount and/or omits at least one ingredientpresent in the prior art product or composition.

1. A chewing gum comprising: one or more allergens; a gum base; one ormore flavoring agents; and a sweetening agent.
 2. The chewing gumaccording to claim 1, further comprising one or more additionalingredients selected from humectants, softeners, coloring agents,stabilizers, or preservatives.
 3. The chewing gum according to claim 1,wherein the gum base is selected from chicle, a natural latex product, anatural gum, or a synthetic rubber.
 4. The chewing gum according toclaim 1, wherein the one or more allergens is selected from a plantpollen allergen, a dust allergen, an animal dander allergen, a moldallergen, a food allergen, and a venom allergen.
 5. The chewing gumaccording to claim 1, wherein the one or more allergens is provided in aform selected from a natural form, an extract, or a recombinantlyproduced form.
 6. The chewing gum according to claim 1, wherein the oneor more allergens is a protein.
 7. The chewing gum according to claim 1,wherein the one or more allergens comprises an environmental allergenselected to correspond with an allergic response of a subject.
 8. Thechewing gum according to claim 1, wherein the chewing gum is acenter-filled chewing gum comprising a soft or liquid center, optionallywherein the soft or liquid center is surrounded at least in part by abarrier layer.
 9. The chewing gum according to claim 8, wherein the oneor more allergens is present in the soft or liquid center of thecenter-filled chewing gum.
 10. The chewing gum according to claim 1,wherein the chewing gum is provided in a unit form selected from astick, a ribbon, or a ball.
 11. A method of decreasing sensitivity toone or more allergens in a subject in need thereof, the methodcomprising the subject chewing a chewing gum comprising: one or moreallergens selected to correspond with an allergic response of thesubject; a gum base; one or more flavoring agents; a sweetening agent;and one or more additional ingredients selected from humectants,softeners, coloring agents, stabilizers, or preservatives.
 12. Themethod according to claim 11, wherein the subject chews the chewing gumaccording to a frequency of: 1 to 5 units of chewing gum daily; or 1 to10 units of chewing gum weekly.
 13. The method according to claim 12,wherein the subject begins a regimen of chewing the chewing gum: atleast 7 days; at least 1 to 4 weeks; at least 4 to 8 weeks; at least 8to 16 weeks; at least 16 to 24 weeks; at least 24 to 36 weeks; or atleast 36 to 52 weeks prior to an anticipated exposure to the one or moreallergens.
 14. The method according to claim 12, wherein chewing thechewing gum results in contact of the one or more allergens with thevestibular oral mucosa of the subject.
 15. A method for reducingsymptoms of allergy in a subject in need thereof, the method comprisingthe subject chewing a chewing gum comprising: one or more allergensselected to correspond with an allergic response of the subject; a gumbase; one or more flavoring agents; a sweetening agent; and one or moreadditional ingredients selected from humectants, softeners, coloringagents, stabilizers, or preservatives.
 16. The method according to claim15, wherein the subject chews the chewing gum according to a frequencyof: 1 to 5 units of chewing gum daily; or 1 to 10 units of chewing gumweekly.
 17. The method according to claim 16, wherein the subject beginsa regimen of chewing the chewing gum: at least 7 days; at least 1 to 4weeks; at least 4 to 8 weeks; at least 8 to 16 weeks; at least 16 to 24weeks; at least 24 to 36 weeks; or at least 36 to 52 weeks prior to ananticipated exposure to the one or more allergens.
 18. The methodaccording to claim 16, wherein chewing the chewing gum results incontact of the one or more allergens with the vestibular oral mucosa ofthe subject.